生物学杂志

生物学杂志 ›› 2023, Vol. 40 ›› Issue (3): 107-.doi: 10.3969/j.issn.2095-1736.2023.03.107

• 技术方法 • 上一篇    下一篇

绿色分离纯化生产高纯度维生素K2

马国良1,2， 郑之明1， 王　鹏1， 王　丽1,2， 赵根海1， 王　晗1   

  1. 1. 中国科学院合肥物质科学研究院智能机械研究所， 合肥 230031;
    2. 中国科学技术大学 研究生院科学岛分院， 合肥 230026
  • 出版日期:2023-06-18 发布日期:2023-06-19
  • 通讯作者: 王晗，助理研究员，主要从事生物化工研究，E-mail:wanghan@ipp.ac.cn；郑之明，研究员，主要从事生物化工研究，E-mail:zhengzhiming2014@163.com；王晗和郑之明为共同通信作者
  • 作者简介:马国良，硕士研究生，主要研究方向为生物工程，E-mail:282010471@qq.com
  • 基金资助:
    安徽省重点研究与开发计划：“针对骨质疏松症的维生素K2制剂开发和应用研究”（No.202004b11020014）; 国家重点研发计划合成生物学专项：“智能工业菌株高效合成技术体系” (No.2019YFA0904300, No.2019YFA0904304)； 安徽省科技重大专项(No.202103a06020003)； 安徽省重点研发计划项目(No.1804b06020342)； 安徽省自然科学基金项目(No.1908085MB48,No.1908085MB43)

Green separation and purification of high purity vitamin K2

MA Guoliang1,2, ZHENG Zhiming1, WANG Peng1, WANG Li1,2, ZHAO Genhai1, WANG Han1   

  1. 1. Institute of Intelligent Machinery, Hefei Institutes of Physical Science, Chinese Academy of Sciences,
    Hefei 230031, China; 2. Science Island Branch of Graduate School, University of Science and Technology of
    China, Hefei 230026, China
  • Online:2023-06-18 Published:2023-06-19

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摘要： 为克服维生素K2（Vitamin K2, VK2）提取工艺有毒溶剂用量大、有毒物质残留等缺点，建立一种高纯度VK2绿色分离纯化工艺方法。使用天然油剂与甲醇、乙醇等有机试剂分别萃取纳豆芽孢杆菌干菌体内的VK2，结果表明，中链甘油三酯的萃取效果最好，最大提取量为2.7 mg/g。通过使用7种不同型号的大孔树脂分离中链甘油三酯溶剂相中的VK2，以及利用静态吸附与解吸附实验，筛选出最优型大孔树脂HC-200S。进一步优化大孔树脂的动态吸附和解吸附工艺，并且绘制大孔树脂的动态解吸附曲线，结果表明：上样液吸附流速为2.0 mL/min、解吸剂为无水乙醇/乙酸丁酯(1∶2,体积比）的混合溶液、解吸剂流速为1.0 mL/min时，HC-200S型大孔树脂对VK2的吸附和解吸附效果最佳；VK2纯度较纯化前提升4倍，达到87.8%，回收率达到93.1%。将树脂分离纯化得到的VK2解吸液经过冷冻结晶和溶剂挥发结晶，分别得到纯度为96.7%和97.1%的VK2晶体。研究将为VK2的工业生产和绿色分离纯化提供方法依据，同时也为其他脂溶性维生素的分离纯化提供借鉴。

关键词: font-size:9.00pt, line-height:150%, ">font-size:9.00pt, line-height:150%, ">维生素K2； 大孔树脂； 分离纯化； 绿色提取； MK-7晶体

Abstract: In order to overcome the shortcomings of high purity Vitamin K2 extraction process, such as large amount of toxic solvent, serious pollution and toxic substance residues, a green separation and purification process of high purity Vitamin K2 was established. Natural oil, methanol, ethanol and other organic reagents were used to extract VK2 from Bacillus natto. The results showed that medium chain triglyceride had the best extraction effect, and the maximum extraction amount was 2.7 mg/g. Seven types of macroporous resins were used to separate VK2 from medium chain triglyceride solvent phase, and the optimal macroporous resin HC-200S was selected by static adsorption and desorption experiments. The dynamic adsorption and desorption process of macroporous resin was optimized, and the dynamic desorption curve of macroporous resin was drawn. The results showed that when the adsorption flow rate of sample solution was 2.0 mL/min, the desorption agent was anhydrous ethanol/butyl acetate mixture solution (1∶2, volume ratio) and the flow rate of desorption agent was 1.0 mL/min, the adsorption and desorption effect of HC-200S macroporous resin on VK2 was the best. At this time, the purity of vitamin K2 was 4 times higher than that of the before purification, reaching 87.8%, and the recovery rate reached 93.1%. VK2 desorption solutions obtained by resin separation and purification were frozen crystallization and solvent volatilization crystallization, respectively, to obtain VK2 crystals with purity of 96.7% and 97.1%. This study provided a method basis for the industrial production and green separation and purification of VK2.

Key words: vitamin K2, macroporous resin, separation and purification, green extraction, MK-7 crystal

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